ELISA Detection Diluent

Product overview

An optimal formulation of buffered solution containing mammalian protein and detergent to dilute detection antibodies and HRP conjugates. Mammalian proteins and salt additives in the ELISA Detection Diluent (1x) provide structural support to both the antibody and HRP-conjugate, preserving enzymatic and IgG activity. The mammalian protein and detergent inhibit detection antibody and HRP conjugates from binding to nonspecific sample proteins, capture antibody, and ELISA plate surface, thus reduce non-specific background noise. This buffer is aseptically filtered through a 0.2 micron filter into clean, pre-sterilized containers. It contains a non-azide, non-mercury preservative.

Storage Condition

Can be stored for up to 1 month at 2-8 °C. Protect from moisture and light. No special shipping conditions or precautions are required.

Application

• Dilute the detection primary antibody to the desired concentration and volume in 1x ELISA Detection Diluent, mix well. Typically, detection antibody concentration can range from 0.05 – 0.5 µg/mL
• Add 100 µL of the diluted detection primary antibody solution in each well and incubate according to the ELISA protocol
• After incubation remove solution and wash the plate 3-4 times with 300 µL of (1x) ELISA Wash Buffer and decant properly
• Dilute the secondary antibody-HRP or streptavidin-HRP conjugate (if biotinylated detection primary antibody was used) to the desired concentration and volume in 1x ELISA Detection Diluent.
• Add 100 µL of the diluted HRP conjugate solution in each well and incubate according to the ELISA protocol
• After incubation remove solution and wash the plate 4-5 times with 300 µL of (1x) ELISA Wash Buffer and decant properly to prepare the plate for HRP-substrate addition.